Suppression of HBV replication by the expression of nickase- and nuclease dead-Cas9
Abstract Complete removal of hepatitis B virus (HBV) DNA from nuclei is difficult by the current therapies. Recent reports have shown that a novel genome-editing tool using Cas9 with a single-guide RNA (sgRNA) system can cleave the HBV genome in vitro and in vivo. However, induction of a double-stra...
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Nature Portfolio
2017
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oai:doaj.org-article:6a0a9b8099f94b9897ea32cd37e0cdaf2021-12-02T16:06:55ZSuppression of HBV replication by the expression of nickase- and nuclease dead-Cas910.1038/s41598-017-05905-w2045-2322https://doaj.org/article/6a0a9b8099f94b9897ea32cd37e0cdaf2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05905-whttps://doaj.org/toc/2045-2322Abstract Complete removal of hepatitis B virus (HBV) DNA from nuclei is difficult by the current therapies. Recent reports have shown that a novel genome-editing tool using Cas9 with a single-guide RNA (sgRNA) system can cleave the HBV genome in vitro and in vivo. However, induction of a double-strand break (DSB) on the targeted genome by Cas9 risks undesirable off-target cleavage on the host genome. Nickase-Cas9 cleaves a single strand of DNA, and thereby two sgRNAs are required for inducing DSBs. To avoid Cas9-induced off-target mutagenesis, we examined the effects of the expressions of nickase-Cas9 and nuclease dead Cas9 (d-Cas9) with sgRNAs on HBV replication. The expression of nickase-Cas9 with a pair of sgRNAs cleaved the target HBV genome and suppressed the viral-protein expression and HBV replication in vitro. Moreover, nickase-Cas9 with the sgRNA pair cleaved the targeted HBV genome in mouse liver. Interestingly, d-Cas9 expression with the sgRNAs also suppressed HBV replication in vitro without cleaving the HBV genome. These results suggest the possible use of nickase-Cas9 and d-Cas9 with a pair of sgRNAs for eliminating HBV DNA from the livers of chronic hepatitis B patients with low risk of undesirable off-target mutation on the host genome.Takeshi KuriharaTakasuke FukuharaChikako OnoSatomi YamamotoKentaro UemuraToru OkamotoMasaya SugiyamaDaisuke MotookaShota NakamuraMasato IkawaMasashi MizokamiYoshihiko MaeharaYoshiharu MatsuuraNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017) |
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Medicine R Science Q |
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Medicine R Science Q Takeshi Kurihara Takasuke Fukuhara Chikako Ono Satomi Yamamoto Kentaro Uemura Toru Okamoto Masaya Sugiyama Daisuke Motooka Shota Nakamura Masato Ikawa Masashi Mizokami Yoshihiko Maehara Yoshiharu Matsuura Suppression of HBV replication by the expression of nickase- and nuclease dead-Cas9 |
| description |
Abstract Complete removal of hepatitis B virus (HBV) DNA from nuclei is difficult by the current therapies. Recent reports have shown that a novel genome-editing tool using Cas9 with a single-guide RNA (sgRNA) system can cleave the HBV genome in vitro and in vivo. However, induction of a double-strand break (DSB) on the targeted genome by Cas9 risks undesirable off-target cleavage on the host genome. Nickase-Cas9 cleaves a single strand of DNA, and thereby two sgRNAs are required for inducing DSBs. To avoid Cas9-induced off-target mutagenesis, we examined the effects of the expressions of nickase-Cas9 and nuclease dead Cas9 (d-Cas9) with sgRNAs on HBV replication. The expression of nickase-Cas9 with a pair of sgRNAs cleaved the target HBV genome and suppressed the viral-protein expression and HBV replication in vitro. Moreover, nickase-Cas9 with the sgRNA pair cleaved the targeted HBV genome in mouse liver. Interestingly, d-Cas9 expression with the sgRNAs also suppressed HBV replication in vitro without cleaving the HBV genome. These results suggest the possible use of nickase-Cas9 and d-Cas9 with a pair of sgRNAs for eliminating HBV DNA from the livers of chronic hepatitis B patients with low risk of undesirable off-target mutation on the host genome. |
| format |
article |
| author |
Takeshi Kurihara Takasuke Fukuhara Chikako Ono Satomi Yamamoto Kentaro Uemura Toru Okamoto Masaya Sugiyama Daisuke Motooka Shota Nakamura Masato Ikawa Masashi Mizokami Yoshihiko Maehara Yoshiharu Matsuura |
| author_facet |
Takeshi Kurihara Takasuke Fukuhara Chikako Ono Satomi Yamamoto Kentaro Uemura Toru Okamoto Masaya Sugiyama Daisuke Motooka Shota Nakamura Masato Ikawa Masashi Mizokami Yoshihiko Maehara Yoshiharu Matsuura |
| author_sort |
Takeshi Kurihara |
| title |
Suppression of HBV replication by the expression of nickase- and nuclease dead-Cas9 |
| title_short |
Suppression of HBV replication by the expression of nickase- and nuclease dead-Cas9 |
| title_full |
Suppression of HBV replication by the expression of nickase- and nuclease dead-Cas9 |
| title_fullStr |
Suppression of HBV replication by the expression of nickase- and nuclease dead-Cas9 |
| title_full_unstemmed |
Suppression of HBV replication by the expression of nickase- and nuclease dead-Cas9 |
| title_sort |
suppression of hbv replication by the expression of nickase- and nuclease dead-cas9 |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/6a0a9b8099f94b9897ea32cd37e0cdaf |
| work_keys_str_mv |
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